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Isolation, Culture, and Differentiation of Bone Marrow Stromal Cells and Osteoclast Progenitors from Mice
Published on: January 6, 2018
Total cell pooling in vitro: an effective isolation method for bone marrow-derived multipotent stromal cells.
Ai-Sze Wee1, Chin-Keong Lim, Azhar Mahmood Merican
1Tissue Engineering Group (TEG), National Orthopaedic Centre for Research and Learning (NOCERAL), Department of Orthopaedic Surgery, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
In Vitro Cellular & Developmental Biology. Animal
|May 28, 2013
Summary
A new total cell pooling method for isolating bone marrow-derived multipotent stromal cells (MSCs) is faster and simpler than standard methods. Both techniques yield MSCs with similar proliferation and differentiation capabilities.
Area of Science:
- Cell Biology
- Regenerative Medicine
- Biotechnology
Background:
- Bone marrow-derived multipotent stromal cells (MSCs) possess significant potential for clinical applications due to their in vitro proliferation and multilineage differentiation abilities.
- Current isolation methods for homogeneous MSC populations face challenges regarding their impact on cell proliferation and differentiation capacity.
Purpose of the Study:
- To evaluate a newly established total cell pooling method for isolating rabbit bone marrow-derived MSCs.
- To compare the efficiency, speed, and cost-effectiveness of the total cell pooling method against the standard density gradient centrifugation method.
Main Methods:
- Rabbit bone marrow-derived MSCs were isolated using either the novel total cell pooling method or the standard density gradient centrifugation method.
- Cell isolation time, primary viable cell yield, and population doubling time were assessed.
- Immunophenotypic analysis (CD29, CD44, CD45) and multilineage differentiation assays (chondrogenesis, osteogenesis, adipogenesis) were performed.
Main Results:
- The total cell pooling method was 65 minutes faster than the standard method.
- No significant differences were observed between the two methods in terms of primary viable cell number and population doubling time (p > 0.05).
- Cells isolated by both methods expressed MSC markers (CD29, CD44) but lacked hematopoietic markers (CD45) and demonstrated multilineage differentiation potential.
Conclusions:
- Both the total cell pooling and standard density gradient centrifugation methods are effective for isolating rabbit bone marrow-derived MSCs with similar biological characteristics.
- The total cell pooling method offers advantages in terms of technical simplicity and cost-effectiveness compared to the standard isolation technique.
